Comparative effects of neurally adjusted ventilatory assist and variable pressure support on lung and diaphragmatic function in a model of acute respiratory distress syndrome: A randomised animal study.

BACKGROUND: Variable assisted mechanical ventilation has been shown to improve lung function and reduce lung injury. However, differences between extrinsic and intrinsic variability are unknown. OBJECTIVE: To investigate the effects of neurally adjusted ventilatory assist (NAVA, intrinsic variability), variable pressure support ventilation (Noisy PSV, extrinsic variability) and conventional pressure-controlled ventilation (PCV) on lung and diaphragmatic function and damage in experimental acute respiratory distress syndrome (ARDS). DESIGN: Randomised controlled animal study. SETTING: University Hospital Research Facility. SUBJECTS: A total of 24 juvenile female pigs. INTERVENTIONS: ARDS was induced by repetitive lung lavage and injurious ventilation. Animals were randomly assigned to 24 h of either: 1) NAVA, 2) Noisy PSV or 3) PCV (n=8 per group). Mechanical ventilation settings followed the ARDS Network recommendations. MEASUREMENTS: The primary outcome was histological lung damage. Secondary outcomes were respiratory variables and patterns, subject-ventilator asynchrony (SVA), pulmonary and diaphragmatic biomarkers, as well as diaphragmatic muscle atrophy and myosin isotypes. RESULTS: Global alveolar damage did not differ between groups, but NAVA resulted in less interstitial oedema in dorsal lung regions than Noisy PSV. Gas exchange and SVA incidence did not differ between groups. Compared with Noisy PSV, NAVA generated higher coefficients of variation of tidal volume and respiratory rate. During NAVA, only 40.4% of breaths were triggered by the electrical diaphragm signal. The IL-8 concentration in lung tissue was lower after NAVA compared with PCV and Noisy PSV, whereas Noisy PSV yielded lower type III procollagen mRNA expression than NAVA and PCV. Diaphragmatic muscle fibre diameters were smaller after PCV compared with assisted modes, whereas expression of myosin isotypes did not differ between groups. CONCLUSION: Noisy PSV and NAVA did not reduce global lung injury compared with PCV but affected different biomarkers and attenuated diaphragmatic atrophy. NAVA increased the respiratory variability; however, NAVA yielded a similar SVA incidence as Noisy PSV. TRIAL REGISTRATION: This trial was registered and approved by the Landesdirektion Dresden, Germany (AZ 24-9168.11-1/2012-2).

The Obliteration of Noncritical Size Bone Defects With Bone Dust or Bone Replacement Material (Bioactive Glass S53P4).

HYPOTHESIS: Bone dust (BD) harvested during operation may be suitable as an autologous obliteration material for noncritical size defects. Bioactive glass (BA) can be an alternative. BACKGROUND: To treat noncritical size defects, BD and BA are commonly used for obliteration techniques. However, the optimal harvesting method and parameters for BD have not been examined. In this study, we analyzed the osseoregenerative potential of both materials. METHODS: Thirteen female merino sheep (7-yr old) underwent surgery on the frontal calvaria. Three defects were inserted. The first defect was considered a reference and remained unfilled, the second defect was filled with BD from the calvaria bone, and the third defect was filled with BA S53P4. The animals were sacrificed after 3 weeks. To evaluate bone regeneration, we used digital volume tomography, bone density measurement, fluorochrome sequence labeling, and histological analysis. RESULTS: All analyses showed quantitative and qualitative bone regeneration 3 weeks after operation. The control blank defect showed significantly less new bone growth than the BD-filled defect. Moreover, bone regeneration occurred from the surrounding bone and showed only a defect bridge in the BD-filled defect. The BA completely filled the defect and had the highest density although the same amount of new mineralized bone generated as in the reference. CONCLUSION: BD and BA seemed to be suitable bone replacement materials for obliteration techniques because they completely filled the defects. Thus, BD harvested under standardized conditions provided a higher level of osteoreparation potential for the generation of woven bone and establishment of defect bridges.

Expression rate of myogenic regulatory factors and muscle growth factor after botulinum toxin A injection in the right masseter muscle of dystrophin deficient (mdx) mice.

BACKGROUND: The mdx mouse, the most approved animal model for basic research in Duchenne muscular dystrophy (DMD), has the ability to compensate muscle degeneration by regeneration process, which is obvious at approx. 3 months of age. Hence, this mouse model is only temporarily suitable to proof craniofacial changes which are usually evident in humans with the progression of the disease. OBJECTIVES: The purpose of our study was to examine the impact of botulinum toxin A (BTX-A) in influencing muscle regeneration in the masticatory muscles of healthy and mdx mice. MATERIAL AND METHODS: Chemo-denervation of the right masseter muscle was induced in 100-day-old, healthy and dystrophic mice by a specific intramuscular BTX-A injection. Gene expression and protein content of myogenic regulatory factors and muscle growth factor (MyoD1, myogenin and myostatin) in the right and left masseter, temporal and the tongue muscle were determined 4 and 21 days after injection, respectively, using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot technique. RESULTS: The 4 day and 21 day interval proved significant but varying changes of mRNA expression in both control and mdx mice. At the protein level, myogenin expression was increased in the temporal and masseter muscle on the injection side in controls, whereas dystrophic mice showed the same effect for MyoD1 expression. Additionally, increased protein expression of all studied genes could be found in dystrophic mice compared to controls, except the left temporal and the tongue muscle. CONCLUSIONS: Muscle regeneration is not constant in BTX-A injected mdx masticatory muscles, presumably due to the already exhausted capacity or functional loss of satellite cells caused by dystrophin deficiency, and, therefore, disturbed regeneration potential of myofibrils. Botulinum toxin A injection cannot fully break down regulatory processes at molecular level in 100-day-old mdx mice. Further investigations are necessary to fully understand the regeneration process following BTX-A injection into dystrophic muscles.

Histological examinations of the in vivo biocompatibility of oxycellulose implanted into rat skeletal muscle.

BACKGROUND: Recently it was shown that oxycellulose suppressed bone regeneration led to an accumulation of connective tissue as well as foam cells in bone defects. OBJECTIVES: Since oxycellulose can be used as a hemostatic agent in general and dental surgery, the aim of the study was to examine muscle tissue response to implanted oxidized cellulose. MATERIAL AND METHODS: RESO-Cell® (Resorba Wundversorgung GmbH, Nuremberg, Germany) standard was implanted in the latissimus dorsi of 20 rats; subsequently, 12 samples were processed for histological evaluation after 4 and 8 weeks. The remaining 8 samples were processed for mRNA expression analyses of gene-encoding growth factors and collagens using quantitative reverse transcription polymerase chain reaction (RT-qPCR). RESULTS: Muscle tissue exposed to oxycellulose showed elevated mRNA levels of COL1A1 compared to untreated muscle tissue. The histological analysis revealed that no undegraded oxycellulose was detectable after as little as 4 weeks. Furthermore, a strong accumulation of CD68-positive foam cells was found in the treated area. CONCLUSIONS: In conclusion, the study has shown that oxidized cellulose can cause an inflammatory response after this material is implanted in skeletal muscle. Therefore, it is not recommended to leave this material in the body over a long period. However, it could be used as auxiliary material in the treatment of periodontal defects.

Histological comparison between laser microtome sections and ground specimens of implant-containing tissues.

Evaluation of bone regeneration and peri-implant bone apposition can only be accomplished using laboratory techniques that allow assessment of decalcified hard tissue. It is known that 5-15µm thick sections can be prepared with the cutting-grinding technique, but their production causes a high material loss (≥0.5mm) between two sections and requires years of training and experience. With the development of the laser microtome it has become possible to cut decalcified bone without high sample material loss. Many scientific publications deal with the application possibilities of the individual methods So far, there is no comparison work between the cutting-grinding technique and laser microtomy. For this reason, new tissue sections were prepared by laser microtome and analyzed histologically from samples that had been previously been prepared by the cutting-grinding technique. Using both methods, it could be demonstrated that the different implants were completely surrounded by a connective tissue layer. In sections (50-100µm) produced by the routine cutting-grinding technique, magnifications up to 20× revealed no detailed histological information because cell structures could not be clearly identified. By contrast, laser microtome sections (10µm) revealed these information as e.g. osteocytes are already clearly visible at 10× magnification. Furthermore, the interface between implant and the surrounding bone could be clearly demonstrated due to visible demarcation between a capsule and connective tissue. At the histological level, laser microtome sections were clearly superior at thicknesses ≥30µm compared to sections produced by the cutting-grinding technique. In addition, laser microtomy has the advantages of time saving and markedly reduced sample loss, especially in cases of the production of serial sections.

The Influence of the Crown-Implant Ratio on the Crestal Bone Level and Implant Secondary Stability: 36-Month Clinical Study.

INTRODUCTION: When the era of dental implantology began, the pioneers defined some gold standards used in dental prosthetics treatment for implant-supported restorations. Referring to traditional prosthetics, it was taken for granted that the length of an implant placed in the alveolar bone (the equivalent of the root) should exceed the length of the superstructure. AIM OF THE STUDY: The aim of the study was to determine whether implant length and the crown-to-implant (C/I) ratio influence implant stability and the loss of the surrounding marginal bone and whether short implants can be used instead of sinus augmentation procedures. MATERIAL AND METHODS: The patients participating in the study (n = 30) had one single tooth implant, a short (OsseoSpeed™ L6 Ø4 mm, Implants) or a regular implant (OsseoSpeed L11 and L13 Ø4 mm, DENTSPLY Implants), placed in the maxilla. The evaluation was based on clinical and radiological examination. The crown-to-implant ratio was determined by dividing the length of the crown together with the abutment by the length of the implant placed crestally. Mean crown-to-implant ratios were calculated separately for each group and its correlation with the MBL (marginal bone loss) and stability was assessed. The authors compared the correlation between the C/I ratio values, MBL, and secondary implant stability. RESULTS: Positive results in terms of primary and secondary stability were achieved with both (short and conventional) implants. The MBL was low for short and conventional implants being 0.34 ± 0.24 mm and 0.22 ± 0.46 mm, respectively. No significant correlation was found between the C/I ratio and secondary stability as well as the C/I ratio and the marginal bone loss. CONCLUSIONS: Short implants can be successfully used to support single crowns. The study has revealed no significant differences in the clinical performance of prosthetic restorations supported by short implants. Clinical trial registration number is NCT03471000.

New nano-hydroxyapatite in bone defect regeneration: A histological study in rats.

Many types of bone substitute materials are available on the market. Researchers are refining new bone substitutes to make them comparable to autologous grafting materials in treatment of bone defects. The purpose of the study was to evaluate the osseoconductive potential and bone defect regeneration in rat calvaria bone defects treated with new synthetic nano-hydroxyapatite. The study was performed on 30 rats divided into 5 equal groups. New preproduction of experimental nano-hydroxyapatite material by NanoSynHap (Poznan, Poland) was tested and compared with commercially available materials. Five mm critical size defects were created and filled with the following bone grafting materials: 1) Geistlich Bio-Oss®; 2) nano-hydroxyapatite+ß-TCP; 3) nano-hydroxyapatite; 4) nano-hydroxyapatite+collagen membrane. The last group served as controls without any augmentation. Bone samples from calvaria were harvested for histological and micro-ct evaluation after 8 weeks. New bone formation was observed in all groups. Histomorphometric analysis revealed an amount of regenerated bone between 34.2 and 44.4% in treated bone defects, whereas only 13.0% regenerated bone was found in controls. Interestingly, in group 3, no significant particles of the nano-HA material were found. In contrast, residual bone substitute material could be detected in all other test groups. Micro-CT study confirmed the results of the histological examinations. The new nano-hydroxyapatite provides comparable results to other grafts in the field of bone regeneration.

In vivo analysis of covering materials composed of biodegradable polymers enriched with flax fibers.

BACKGROUND: The objective of this study was to investigate the in vivo effect of bioactive composites with poly(lactic acid) (PLA) or polycaprolactone (PCL) as the matrix, reinforced with bioplastic flax fibers, on the surrounding muscle tissue. METHODS: Materials of pure PLA and PCL and their composites with flax fibers from genetically modified plants producing poly-3-hydroxybutyrate (PLA-transgen, PCL-transgen) and unmodified plants (PLA-wt, PCL-wt) were placed subcutaneous on the M. latissimus dorsi for four weeks. RESULTS: The analysis of histological samples revealed that every tested material was differently encapsulated and the capsule thickness is much more pronounced when using the PCL composites in comparison with the PLA composites. The encapsulation by connective tissue was significantly reduced around PCL-transgen and significantly increased in the cases of PLA-transgen and PLA-wt. In the collected muscle samples, the measured protein expression of CD45, lymphocyte common antigen, was significantly increased after the use of all tested materials, with the exception of pure PCL. In contrast, the protein expression of caveolin-1 remained unchanged after treatment with the most examined materials. Only after insertion of PLA-wt, a significant increase of caveolin-1 protein expression was detected, due to the improved neovascularization. CONCLUSION: These data support the presumption that the new bioactive composites are biocompatible and they could be applicable in the medical field to support the regenerative processes.

Application of tissue-engineered bone grafts for alveolar cleft osteoplasty in a rodent model.

OBJECTIVES: The clinical standard for alveolar cleft osteoplasty is augmentation with autologous bone being available in limited amounts and might be associated with donor site morbidity. The aim of the present study was the creation of tissue-engineered bone grafts and their in vivo evaluation regarding their potential to promote osteogenesis in an alveolar cleft model. MATERIALS AND METHODS: Artificial bone defects with a diameter of 3.3 mm were created surgically in the palate of 84 adult Lewis rats. Four experimental groups (n = 21) were examined: bovine hydroxyl apatite/collagen (bHA) without cells, bHA with undifferentiated mesenchymal stromal cells (MSC), bHA with osteogenically differentiated MSC. In a control group, the defect remained empty. After 6, 9 and 12 weeks, the remaining defect volume was assessed by cone beam computed tomography. Histologically, the remaining defect width and percentage of bone formation was quantified. RESULTS: After 12 weeks, the remaining defect width was 60.1% for bHA, 74.7% for bHA with undifferentiated MSC and 81.8% for bHA with osteogenically differentiated MSC. For the control group, the remaining defect width measured 46.2% which was a statistically significant difference (p < 0.001). CONCLUSIONS: The study design was suitable to evaluate tissue-engineered bone grafts prior to a clinical application. In this experimental set-up with the described maxillary defect, no promoting influence on bone formation of bone grafts containing bHA could be confirmed. CLINICAL RELEVANCE: The creation of a sufficient tissue-engineered bone graft for alveolar cleft osteoplasty could preserve patients from donor site morbidity.

Influence of Botulinumtoxin A on the Expression of Adult MyHC Isoforms in the Masticatory Muscles in Dystrophin-Deficient Mice (Mdx-Mice).

The most widespread animal model to investigate Duchenne muscular dystrophy is the mdx-mouse. In contrast to humans, phases of muscle degeneration are replaced by regeneration processes; hence there is only a restricted time slot for research. The aim of the study was to investigate if an intramuscular injection of BTX-A is able to break down muscle regeneration and has direct implications on the gene expression of myosin heavy chains in the corresponding treated and untreated muscles. Therefore, paralysis of the right masseter muscle was induced in adult healthy and dystrophic mice by a specific intramuscular injection of BTX-A. After 21 days the mRNA expression and protein content of MyHC isoforms of the right and left masseter, temporal, and the tongue muscle were determined using quantitative RT-PCR and Western blot technique. MyHC-IIa and MyHC-I-mRNA expression significantly increased in the paralyzed masseter muscle of control-mice, whereas MyHC-IIb and MyHC-IIx/d-mRNA were decreased. In dystrophic muscles no effect of BTX-A could be detected at the level of MyHC. This study suggests that BTX-A injection is a suitable method to simulate DMD-pathogenesis in healthy mice but further investigations are necessary to fully analyse the BTX-A effect and to generate sustained muscular atrophy in mdx-mice.

Bone Regeneration after Treatment with Covering Materials Composed of Flax Fibers and Biodegradable Plastics: A Histological Study in Rats.

The aim of this study was to examine the osteogenic potential of new flax covering materials. Bone defects were created on the skull of forty rats. Materials of pure PLA and PCL and their composites with flax fibers, genetically modified producing PHB (PLA-transgen, PCL-transgen) and unmodified (PLA-wt, PCL-wt), were inserted. The skulls were harvested after four weeks and subjected to histological examination. The percentage of bone regeneration by using PLA was less pronounced than after usage of pure PCL in comparison with controls. After treatment with PCL-transgen, a large amount of new formed bone could be found. In contrast, PCL-wt decreased significantly the bone regeneration, compared to the other tested groups. The bone covers made of pure PLA had substantially less influence on bone regeneration and the bone healing proceeded with a lot of connective tissue, whereas PLA-transgen and PLA-wt showed nearly comparable amount of new formed bone. Regarding the histological data, the hypothesis could be proposed that PCL and its composites have contributed to a higher quantity of the regenerated bone, compared to PLA. The histological studies showed comparable bone regeneration processes after treatment with tested covering materials, as well as in the untreated bone lesions.

A silver carp skin derived collagen in bone defect treatment-A histological study in a rat model.

In recent years, there has been increasing interest in elaboration of novel therapeutic strategies, such as the use of the marine collagen products. Biochemical properties of marine collagen are different from those of mammalian collagen; e.g., its extremely high solubility in diluted acid. Extracts produced using low temperature techniques contain a number of small proteins and collagen with preserved triple helix structure. The aim of the study was to evaluate the influence of a new marine product Collgel® obtained with a unique method from a silver carp (Hypophthalmichthys molitrix) on bone defect healing in a rat study. For this purpose bone defects with diameters of 5mm were created in 15 animals and subsequently filled with Collgel combined with another commercially available material. Samples were processed for histological evaluation and a Micro-CT study was performed. Histological analysis showed new bone formation in all groups after 8 weeks. The bone formation was significantly increased in treated bone lesions compared to untreated bone tissue. However no significant difference was noted between the healing of the defects filled with xenogenic bovine derived bone substitute alone and xenogenic, bovine derived bone substitute combined with a marine delivered collagen. Finding from the histological examination was confirmed in a Micro-CT study. The study has shown that the new marine product can be used instead of conventional porcine or bovine collagen membranes in guided bone regeneration.

Interrelationship between bone substitution materials and skeletal muscle tissue.

Bone density and quantity are primary conditions for the insertion and stability of dental implants. In cases of a lack of adequate maxillary or mandibulary bone, bone augmentation will be necessary. The use of synthetic bioactive bone substitution materials is of increasing importance as alternatives to autogenously bone grafts. It is well known that bone can influence muscle function and muscle function can influence bone structures. Muscles have a considerable potential of adaptation and muscle tissue surrounding an inserted implant or bone surrogate can integrate changes in mechanical load of the muscle and hereupon induce signaling cascades with protein synthesis and arrangement of the cytoskeleton. The Musculus latissimus dorsi is very often used for the analyses of the in vivo biocompatibility of newly designed biomaterials. Beside macroscopically and histologically examination, biocompatibility can be assessed by analyses of the biomaterial influence of gene expression. This review discusses changes in the fiber type distribution, myosin heavy chain isoform composition, histological appearance and vascularization of the skeletal muscle after implantation of bone substitution materials. Especially, the effects of bone surrogates should be described at the molecular-biological and cellular level.

Socket augmentation using a commercial collagen-based product--an animal study in pigs.

The aim of the present study was to identify properties of pure collagen for augmentation techniques and compare to a proved xenogenic material and natural bone regeneration. For that the osteogenesis of extraction alveoli after augmentation with a collagen cone covered with an absorbable collagen membrane in a single product (PARASORB Sombrero®, Resorba) was evaluated in a pig model. Extraction alveoli were treated with the collagen cone and the collagen membrane in a single product (test group; n=7) or demineralized bovine bone mineral and a collagen membrane (two separate products; positive control; n=7). Untreated alveoli were used (n=6) as negative controls.(1) Bone specimens were extracted 1 and 3 months after teeth extraction. Serial longitudinal sections were stained with Masson Goldner trichrome. Furthermore, bone specimens were examined using X-ray analyses. Significant differences of bone atrophy were detected 12 weeks after material insertion using X-ray analyses. The bone atrophy was reduced by approximately 32% after insertion of the positive control (P=0.046). Bone atrophy reached 37.6% of those from untreated alveoli (P=0.002) using the test group. After 4 weeks, bone formation was noticeable in most sites, whereas after 12 weeks of healing, specimens of all groups exhibited nearly complete osseous organization of the former defected area. The mandibulary bone texture showed typical spongious bone structures. Histomorphometric analyses revealed after 4 and 12 weeks significant higher levels of bone marrow in test and negative control than in positive control. Quantification of bone tissue and osteoid does not show any significant difference. The present study confirms reduced bone resorption following socket augmentation with an absorbable collagen membrane with collagen cone while the resulting bone structure is similar to natural bone regeneration. Pure collagen can be used for bone augmentation, and shows over other xenogenic materials, a clear advantage with respect to the bone density and structure.

Comparative study of biphasic calcium phosphate with beta-tricalcium phosphate in rat cranial defects--A molecular-biological and histological study.

The aim of this study was to evaluate the in vivo biocompatibility of a biphasic calcium phosphate (BCP) bone graft substitute consisting of 60% hydroxyapatite and 40% ß-tricalcium phosphate (ß-TCP) in comparison to a pure ß-TCP of identical shape and porosity. The materials were evaluated using an established rat cranial defect model in 24 animals. One bone defect with a diameter of 5mm was created per animal. The defects were filled with either BCP or ß-TCP and left to heal for 4 weeks. Twelve samples (6 per material) were processed for histological evaluation and immunohistochemistry. The remaining 12 samples were processed for mRNA expression analysis. No signs of inflammation or adverse material reactions were detected. New bone formation in the former defect site did not differ between the two groups (BCP: 49.2%; ß-TCP: 52.4%). Osteoblast-like and TRAP-positive osteoclast-like cells were found at the surface of the bone graft substitute granules. The ß-TCP group showed significantly higher mRNA levels for the bone resorption marker Acp5 and osteogenic differentiation marker Runx2. The expression of IGF1, IGF2, VEGF, Phex, Alpl, Col1, Col2, Bglap and MMP8 did not differ between the groups. The in vivo biocompatibility of BCP is to a large part identical to those of TCP. Within the limitation of the animal model, the implantation study shows that BCP can be used as bone graft substitute, due to the fact that the material integrates into tissue, remains stable in the implantation bed and serves as an osteoconductive scaffold.

Histological and molecular-biological analyses of poly(3-hydroxybutyrate) (PHB) patches for enhancement of bone regeneration.

Tissue engineered cell-seeded constructs with poly(3)hydroxybutyrate (PHB) induced ectopic bone formation after implantation into the back muscle of rats. The objective of our in vivo study was to evaluate the osteogenic potential of pure PHB patches in surgically created cranial defects. For this, PHB patches were analyzed after implantation in surgically created defects on the cranium of adult male rats. After healing periods of 4, 8 and 12 weeks, the bone tissue specimens containing PHB patches were processed and analyzed histologically as well as molecular-biologically. After 4 weeks, the PHB patches were completely embedded in connective tissue. Eight weeks after PHB insertion, bone regeneration proceeding from bearing bone was found in 50% of all treated animals, whereas all PHB treated cavities showed both bone formation and embedding of the patches in bone 12 weeks after surgery. Furthermore, all slices showed pronounced development of blood vessels. Histomorphometric analysis presented a regenerated bone mean value between 46.4 ± 16.1% and 54.2 ± 19.3% after 4-12 weeks of healing. Caveolin-1 staining in capillary-like structures showed a 1.16-1.38 fold increased expression in PHB treated defects compared to controls. Real-time RT-PCR analyses showed significantly lower expressions of Alpl, Col1a1 and VEGFA in cranium defects after treatment with PHB patches compared to untreated bony defects of the same cranium. Within the limits of the presented animal investigation, it could conclude that the tested PHB patches featured a good biocompatibility and an osteoconductive character.